Genetic and chemical rescue of the Saccharomyces cerevisiae phenotype induced by mitochondrial DNA polymerase mutations associated with progressive external ophthalmoplegia in humans

The human POLG gene encodes the catalytic subunit of mitochondrial DNA polymerase γ (pol γ). Mutations in pol γ are associated with a spectrum of disease phenotypes including autosomal dominant and recessive forms of progressive external ophthalmoplegia, spino-cerebellar ataxia and epilepsy, and Alpers-Huttenlocher hepatocerebral poliodystrophy. Multiple deletions, or depletion of mtDNA in affected tissues, are the molecular hallmarks of pol γ mutations. To shed light on the pathogenic mechanisms leading to these phenotypes, we have introduced in MIP1, the yeast homologue of POLG, two mutations equivalent to the human Y955C and G268A mutations, which are associated with dominant and recessive PEO, respectively. Both mutations induced the generation of petite colonies, carrying either rearranged (ρ−) or no (ρ0) mtDNA. Mutations in genes that control the mitochondrial supply of deoxynucleotides (dNTP) affect the mtDNA integrity in both humans and yeast. To test whether the manipulation of the dNTP pool can modify the effects of pol γ mutations in yeast, we have overexpressed a dNTP checkpoint enzyme, ribonucleotide reductase, RNR1, or deleted its inhibitor, SML1. In both mutant strains, the petite mutability was dramatically reduced. The same result was obtained by exposing the mutant strains to dihydrolipoic acid, an anti-oxidant agent. Therefore, an increase of the mitochondrial dNTP pool and/or a decrease of reactive oxygen species can prevent the mtDNA damage induced by pol γ mutations in yeast and, possibly, in human

Difficoltà di apprendimento: il ruolo dell’attività motoria finalizzata

A set of motor and sequential skills must be automatized in order to carry out the task of learning. Indeed, the learning process requires the visual search of number/letter shapes, the analysis of left-to-right sequences, memorization of letters shape tracing and eye muscle coordination in reading and writing. The low efficiency of some of the key functions within motor scheme, such as lateralization and eye-hand coordination, negatively affects the organizationof reading-writing and calculation processes. The aim of our study was the assessment of improvements within reading/writing/ calculation outcomes, in LSD children after a specific motor sequence training. In particular, skill coordination performance leaded all children to an important improvements during specific trials about the scholastic practice.Specific and ad-personam sequence of exercise could be an elective teaching methodology to over-stimolate the neuro-motor viability.La forma di numeri e lettere, la loro successione da sinistra a destra che struttura parole e frasi, la memorizzazione del tracciato dei diversi caratteri, la coordinazione della muscolatura oculare utilizzata negli atti dello scrivere e del leggere sono tutti elementi di qualità motoria e sequenziale che l’individuo deve automatizzare per affrontare il compito di apprendere.La scarsa efficienza di alcune delle funzioni chiave della motricità, come la lateralizzazione e la coordinazione oculo manuale, incide pesantemente nell’organizzazione delle procedure della letto-scrittura e del calcolo. Il nostro studio si pone lo scopo di esemplificare come l’abilitazione coordinativa possa attenuare significativamente i DSA attraverso interventi di motricità finalizzata. Il movimento umano inteso come “atto in funzione di uno scopo” e proposto con una didattica protetta e di gruppo si è dimostrata essere in grado diripristinare una viabilità neuro motoria più funzionale a raggiungere il successo nelle pratiche scolastiche

DAS: a data management system for instrument tests and operations

The Data Access System (DAS) is a metadata and data management software system, providing a reusable solution for the storage of data acquired both from telescopes and auxiliary data sources during the instrument development phases and operations. It is part of the Customizable Instrument WorkStation system (CIWS-FW), a framework for the storage, processing and quick-look at the data acquired from scientific instruments. The DAS provides a data access layer mainly targeted to software applications: quick-look displays, pre-processing pipelines and scientific workflows. It is logically organized in three main components: an intuitive and compact Data Definition Language (DAS DDL) in XML format, aimed for user-defined data types; an Application Programming Interface (DAS API), automatically adding classes and methods supporting the DDL data types, and providing an object-oriented query language; a data management component, which maps the metadata of the DDL data types in a relational Data Base Management System (DBMS), and stores the data in a shared (network) file system. With the DAS DDL, developers define the data model for a particular project, specifying for each data type the metadata attributes, the data format and layout (if applicable), and named references to related or aggregated data types. Together with the DDL user-defined data types, the DAS API acts as the only interface to store, query and retrieve the metadata and data in the DAS system, providing both an abstract interface and a data model specific one in C, C++ and Python. The mapping of metadata in the back-end database is automatic and supports several relational DBMSs, including MySQL, Oracle and PostgreSQL.Comment: Accepted for pubblication on ADASS Conference Serie

CIWS-FW: a Customizable InstrumentWorkstation Software Framework for instrument-independent data handling

The CIWS-FW is aimed at providing a common and standard solution for the storage, processing and quick look at the data acquired from scientific instruments for astrophysics. The target system is the instrument workstation either in the context of the Electrical Ground Support Equipment for space-borne experiments, or in the context of the data acquisition system for instrumentation. The CIWS-FW core includes software developed by team members for previous experiments and provides new components and tools that improve the software reusability, configurability and extensibility attributes. The CIWS-FW mainly consists of two packages: the data processing system and the data access system. The former provides the software components and libraries to support the data acquisition, transformation, display and storage in near real time of either a data packet stream and/or a sequence of data files generated by the instrument. The latter is a meta-data and data management system, providing a reusable solution for the archiving and retrieval of the acquired data. A built-in operator GUI allows to control and configure the IW. In addition, the framework provides mechanisms for system error and logging handling. A web portal provides the access to the CIWS-FW documentation, software repository and bug tracking tools for CIWS-FW developers. We will describe the CIWS-FW architecture and summarize the project status.Comment: Accepted for pubblication on ADASS Conference Serie

Vrtlarski rad i izloženost teškim metalima u gradskom okružju

Urban soil may be a source of occupational exposure to various pollutants in gardening and land cultivation. This paper presents data of a one-year follow-up of lead, cadmium, nickel, chromium, and vanadium in the environment of the city of Bologna. Samples of soil and leaves were collected at three locations; gardens from the inner-city high-traffic area, parks in moderatetraffic area, and parks in suburban, low-traffic area. The top and deeper layers of soil and leaves were mainly polluted by lead at all locations, which corresponded to the traffic density. Personal samplers recorded greater concentrations of airborne metals than did the area samplers but the values kept below the threshold limit established by the American Conference of Governmental Industrial Hygienists for the working environment. Due to cumulative nature and interactive effects of toxic metals with other toxic and essential elements, long-term exposure to metals in the urban environment may be a health risk for occupationally exposed gardeners.Gradsko tlo može biti izvorom profesionalne izloženosti različitim onečišćivačima u vrtlara, uključujući otrovne teške metale koji se talože iz onečišćenog zraka na tlo i lišće. U radu su prikazani podaci jednogodišnjeg praćenja koncentracija olova, kadmija, niklja, kroma i vanadija u talijanskom gradu Bologni. Uzorci tla i lišća skupljani su na tri mjesta s različitom gustoćom prometa; u središtu grada s velikom gustoćom prometa, u parkovima s umjerenim prometom u okolišu i u prigradskim parkovima sa slabim prometom. Na svim lokacijama gornji i dublji slojevi tla bili su podjednako onečišćeni ponajprije olovom i to je bilo u svezi s prometnom gustoćom. Koncentracije metala u zraku utvr|ene u filtrima osobnih skupljača bile su više od koncentracija u skupljačima na tlu. Vrijednosti metala u zraku bile su niže od graničnih vrijednosti utvr|enih za radni okoliš od Američke konferencije vladinih industrijskih higijeničara (American Conference of Governmental Industrial Hygienists). Zaključeno je da vrtlari, u usporedbi s općim stanovništvom, i pri niskim razinama izloženosti u gradskom okolišu imaju povećan zdravstveni rizik za štetna djelovanja otrovnih metala i drugih onečišćivača okoliša zbog dugotrajne izloženosti u svezi s njihovim radom

Detection Analysis and Study of Genomic Region Variability of JCPyV, BKPyV, MCPyV, HPyV6, HPyV7 and QPyV in the Urine and Plasma of HIV-1-Infected Patients

Since it was clearly established that HIV/AIDS predisposes to the infection, persistence or reactivation of latent viruses, the prevalence of human polyomaviruses (HPyVs) among HIV-1-infected patients and a possible correlation between HPyVs and HIV sero-status were investigated. PCR was performed to detect and quantify JCPyV, BKPyV, MCPyV, HPyV6, HPyV7 and QPyV DNA in the urine and plasma samples of 103 HIV-1-infected patients. Subsequently, NCCR, VP1 and MCPyV LT sequences were examined. In addition, for MCPyV, the expression of transcripts for the LT gene was investigated. JCPyV, BKPyV and MCPyV’s presence was reported, whereas HPyV6, HPyV7 and QPyV were not detected in any sample. Co-infection patterns of JCPyV, BKPyV and MCPyV were found. Archetype-like NCCRs were observed with some point mutations in plasma samples positive for JCPyV and BKPyV. The VP1 region was found to be highly conserved among these subjects. LT did not show mutations causing stop codons, and LT transcripts were expressed in MCPyV positive samples. A significant correlation between HPyVs’ detection and a low level of CD4+ was reported. In conclusion, HPyV6, HPyV7 and QPyV seem to not have a clinical relevance in HIV-1 patients, whereas further studies are warranted to define the clinical importance of JCPyV, BKPyV and MCPyV DNA detection in these subjects

Resistance to Ceftazidime/Avibactam in Klebsiella pneumoniae KPC-Producing Isolates: A Real-Life Observational Study

Background: Ceftazidime/avibactam (CAZ-AVI) resistance amongst Enterobacterales is worryingly increasing worldwide. Objectives: The aim of this study was to collect and describe real-life data on CAZ-AVI-resistant Klebsiella pneumoniae (KP) isolates in our University Hospital, with the ultimate goal of evaluating possible risk factors related to the acquisition of resistance. Methods: This is a retrospective observational study, including unique Klebsiella pneumoniae (KP) isolates resistant to CAZ-AVI (CAZ-AVI-R) and producing only KPC, collected from July 2019 to August 2021 at Policlinico Tor Vergata, Rome, Italy. The pathogen's list was obtained from the microbiology laboratory; clinical charts of the corresponding patients were reviewed to collect demographic and clinical data. Subjects treated as outpatients or hospitalized for <48 h were excluded. Patients were then divided into two groups: S group, if they had a prior isolate of CAZ-AVI-susceptible KP-KPC, and R group, if the first documented isolate of KP-KPC was resistant to CAZ-AVI. Results: Forty-six unique isolates corresponding to 46 patients were included in the study. The majority of patients (60.9%) were hospitalized in an intensive care unit, 32.6% in internal medicine wards and 6.5% in surgical wards. A total of 15 (32.6%) isolates were collected from rectal swabs, representing a colonization. Amongst clinically relevant infections, pneumonia and urinary tract infections were the most commonly found (5/46, 10.9% each). Half of the patients received CAZ-AVI prior to isolation of the KP-KPC CAZ-AVI-R (23/46). This percentage was significantly higher in patients in the S group compared to patients in the R group (69.3% S group vs. 25% R group, p = 0.003). No differences between the two groups were documented in the use of renal replacement therapy or in the infection site. The clinically relevant CAZ-AVI-R KP infections (22/46, 47.8%) were all treated with a combination therapy, 65% including colistin and 55% including CAZ-AVI, with an overall clinical success of 38.1%. Conclusions: Prior use of CAZ-AVI was associated with the emergence of drug resistance

Assessment of distribution of CD34 epitope classes in fresh and cryopreserved peripheral blood progenitor cells and acute myeloid leukemic blasts.

So far several reports have described changes in the expression of surface antigens in progenitor cells and blasts following cryopreservation. However, there are no data on the effects of cryopreservation on the expression of the three CD34 epitope classes, and on their relationship with the clonogenic capacity of PBPC collected by leukapheresis. DESIGN AND METHODS: In order to analyze the effects of freezing/thawing procedures (Eth 80C storage for 3 months) and use of dimethylsulfoxide (DMSO) on the immunophenotype profile and colony production of peripheral blood progenitor cells (PBPC) in apheresis products derived from 20 patients with stage 0-III non-Hodgkin's lymphoma (nHL), a flow cytometry study was undertaken using different CD34 monoclonal antibodies (MoAbs) capable of recognizing the 3 epitope classes of CD34 molecule (class III: HPCA-2/FITC, HPCA-2/PE, 581/FITC, 581/PE; class II: Q-Bend 10/PE; class I: ICH3/PE, BI3C5-PE, Immu-133-PE). CD34 epitope expression was also analyzed in thawed CD34+ blasts obtained from 14 patients with acute myeloid leukemia (AML), who were analyzed using a larger number (#17) of CD34 epitope class I, II, and III reactive MoAbs. RESULTS: Under our experimental conditions it was found that class III and class II CD34 epitopes (differentially resistant to enzymatic cleavage with neuraminidase, chymopapain and glycoprotease) are better preserved than class I epitope Eth sensitive to degradation Eth after cell exposure to cryoprotectant DMSO and the freezing- thawing procedures. Results further showed a concomitant decrease in class I CD34+ counts and in BFU-E colony production. A significant increase in CD34 antigen expression levels (i.e. antibody binding capacity, ABC) by cryopreserved cells stained with CD34 epitope class III, and class II reactive MoAbs was also documented, while no changes after cryopreservation were noted using class I-reactive MoAbs. The slight increase in the percentage of CD34+ cells detected after frozen storage was correlated to a concomitant decrease in the number of more mature myeloid cells (CD15+, CD13+, CD33+). Compared to pre-cryopreservation values, a slight reduction in class I CD34 epitope expression was also found in thawed CD34+ AML blasts. INTERPRETATION AND CONCLUSIONS: As far as the reduction of class I CD34 epitope is concerned, it may be hypothesized that the freezing procedure, use of DMSO, and/or lysis methodology may either damage a CD34 subset, or induce distinct alterations of the CD34 glycoprotein, possibly determining a reduction in their immunoreactivity with some CD34 MoAbs. In conclusion, this study has shown that exposure to the cryoprotectant DMSO and the freezing/thawing procedures modifies the distribution of CD34 epitopes as well as the clonogenic capacity of PBPCs from nHL patients, and CD34+ blasts from AML. These findings need to considered when selecting CD34 MoAbs for enumeration and positive selection of stem/progenitor cells for research and clinical purposes

Human Arboviral Infections in Italy: Past, Current, and Future Challenges

: Arboviruses represent a public health concern in many European countries, including Italy, mostly because they can infect humans, causing potentially severe emergent or re-emergent diseases, with epidemic outbreaks and the introduction of endemic circulation of new species previously confined to tropical and sub-tropical regions. In this review, we summarize the Italian epidemiology of arboviral infection over the past 10 years, describing both endemic and imported arboviral infections, vector distribution, and the influence of climate change on vector ecology. Strengthening surveillance systems at a national and international level is highly recommended to be prepared to face potential threats due to arbovirus diffusion